High dietary threonine extraction by the digestive tract suggests that threonine contributes to maintain gut physiology. In the present study, we evaluated the impact of a low (6.5 g of threonine/kg diet; LT group) or a control well-balanced threonine diet (9.3 g of threonine/kg diet; C group) given to piglets for 2 weeks on ileal permeability and Na+-dependant glucose absorption capacity in Ussing chambers. The paracellular permeability was significantly increased in the ileum of LT compared to C piglets (P=.017). The Na+-dependent glucose absorption capacity showed a nonsignificant increase in the LT piglets. In addition, we analysed ileal gene expression profiles in the LT and C groups using porcine multitissue cDNA microarrays. Compared to the C piglets, the expression of 324 genes was significantly modified in the ileum of the LT piglets: 214 genes were overexpressed (145 annotated) and 110 were down-expressed (79 annotated). Among them, some are involved in immune and defense responses, energy metabolism and protein synthesis. Furthermore, microarray analysis highlights changes in the expression of the gene encoding for the sodium/glucose cotransporter (SGLT1) and of genes involved in the regulation of paracellular permeability (ZO-1, cingulin and myosin light chain kinase). In conclusion, our results indicate that a moderate threonine deficiency affects intestinal functionality. (C) 2010 Elsevier Inc. All rights reserved.